ERK-dependent IL-6 autocrine signaling mediates adaptive resistance to pan-PI3K inhibitor BKM120 in head and neck squamous cell carcinoma.

Hyperactivation of phosphatidylinositol 3-kinase (PI3K) pathway occurs frequently in head and neck squamous cell carcinoma (HNSCC). However, clinical outcomes of targeting the PI3K pathway have been underwhelming. In present study, we investigated the resistant mechanisms and potential combination therapeutic strategy to overcome adaptive resistance to PI3K inhibitor in HNSCC. Treatment of NVP-BKM120, a pan-PI3K inhibitor, led to upregulation of interleukin-6 (IL-6) and subsequent activation of either extracellular signal-regulated kinase (ERK) or signal transducers and activators of transcription 3 (STAT3), causing modest antitumor effects on the growth of HNSCC cells. Blockade of autocrine IL-6 signaling with siRNA or neutralizing antibody for IL-6 receptor (IL-6R) completely abolished NVP-BKM120-induced activation of ERK and STAT3 as well as expression of c-Myc oncogene, which resulted in enhanced sensitivity to NVP-BKM120. Moreover, when compared with a pharmacologic inhibitor or silencing of STAT3, trametinib, a MEK inhibitor, in combination with NVP-BKM120 yielded more potent anti-proliferative effects by inhibiting S phase transition, arresting cells at G0/G1 phase, and downregulating IL-6 and c-Myc expression. Furthermore, as compared with either agent alone, combination of NVP-BKM120 with trametinib or tocilizumab, a humanized anti-IL-6R antibody, significantly suppressed tumor growth in NVP-BKM120-resistant patient-derived tumor xenograft (PDTX) models, which was also confirmed in PDTX-derived cell lines. Collectively, these results suggested that IL-6/ERK signaling is closely involved in adaptive resistance of NVP-BKM120 in HNSCC cells, providing a rationale for a novel combination therapy to overcome resistance to PI3K inhibitors.

Randomised controlled clinical trial of augmentation of the alveolar ridge using recombinant human bone morphogenetic protein 2 with hydroxyapatite and bovine-derived xenografts: comparison of changes in volume.

The aim of this randomised controlled clinical trial was to assess the early efficacy of bone morphogenetic protein-2 with hydroxyapatite granules (BMP-2/hydroxyapatite) on augmentation of the alveolar ridge, by comparing changes in volume with those associated with the use of an inorganic bovine-derived xenograft (BDX). We studied 20 patients who were divided into two groups using a table of random numbers, and BMP-2/hydroxyapatite and BDX were applied accordingly. Computed tomographic (CT) images and panoramic radiographs were obtained immediately after operation and four months later. CT images were reconstructed in three dimensions to measure volumetric changes, and linear measurements were made on panoramic images. The mean (SD) absorption rates for BMP-2/hydroxyapatite and BDX were 13.2 (8.8)% and 13.8 (20.5)%, respectively. While the mean value did not differ significantly between the two materials, the SD was higher in the BDX group than in the BMP-2/hydroxyapatite group. No clinically important complications occurred in either group. We conclude that both BMP-2/hydroxyapatite and BDX were effective in augmenting the alveolar ridge, but BMP-2/hydroxyapatite seemed to be more useful in complicated bone defects.

Activation of the Met kinase confers acquired drug resistance in FGFR-targeted lung cancer therapy.

Aberrant fibroblast growth factor receptor (FGFR) activation/expression is a common feature in lung cancer (LC). In this study, we evaluated the antitumor activity of and the mechanisms underlying acquired resistance to two potent selective FGFR inhibitors, AZD4547 and BAY116387, in LC cell lines. The antitumor activity of AZD4547 and BAY1163877 was screened in 24 LC cell lines, including 5 with FGFR1 amplification. Two cell lines containing FGFR1 amplifications, H1581 and DMS114, were sensitive to FGFR inhibitors (IC50<250 nm). Clones of FGFR1-amplified H1581 cells resistant to AZD4547 or BAY116387 (H1581AR and H1581BR cells, respectively) were established. Receptor tyrosine kinase (RTK) array and immunoblotting analyses showed strong overexpression and activation of Met in H1581AR/BR cells, compared with that in the parental cells. Gene set enrichment analysis against the Kyoto Encyclopedia of Genes and Genomes (KEGG) database showed that cytokine-cytokine receptor interaction pathways were significantly enriched in H1581AR/BR cells, with Met contributing significantly to the core enrichment. Genomic DNA quantitative PCR and fluorescent in situ hybridization analyses showed MET amplification in H1581AR, but not in H1581BR, cells. Met amplification drives acquired resistance to AZD4547 in H1581AR cells by activating ErbB3. Combination treatment with FGFR inhibitors and an anaplastic lymphoma kinase (ALK)/Met inhibitor, crizotinib, or Met-specific short interfering RNA (siRNA) synergistically inhibited cell proliferation in both H1581AR and H1581BR cells. Conversely, ectopic expression of Met in H1581 cells conferred resistance to AZD4547 and BAY1163877. Acquired resistance to FGFR inhibitors not only altered cellular morphology, but also promoted migration and invasion of resistant clones, in part by inducing epithelial-to-mesenchymal transition. Taken together, our data suggest that Met activation is sufficient to bypass dependency on FGFR signaling. Concurrent inhibition of the Met and FGFR pathways may have synergistic clinical benefits when targeting FGFR-dependent LC.

Joint Cooling does not Hinder Athletic Performance during High-intensity Intermittent Exercise.

We examined the effects of ankle and knee joint cooling on 20-m sprint times and maximal vertical jump heights during high-intensity intermittent exercise. 21 healthy collegiate male basketball (n=14) and handball players (n=7) underwent 3 experimental sessions. Each session consisted of four 15-min quarters of high-intensity intermittent exercises including various intensities of 20-m shuttle running and jumping. A 20-min bilateral joint cooling (ankle, knee, or control-no cooling: in a counterbalanced order) was applied before quarters 1 and 3. After joint cooling, no warm-up activity other than the exercise protocol was given. The 20-m sprint times and maximal vertical jump heights in each experimental session were recorded at baseline (prior to quarter-1) and during each quarter. To test joint cooling effects over time, we performed 3×5 mixed model ANOVAs. Neither ankle nor knee joint cooling changed 20-m sprint times (F8,280=1.45; p=0.18) or maximal vertical jump heights (F8,280=0.76; p=0.64). However, a trend was observed in which joint cooling immediately decreased (quarters 1 and 3) but active warm-up for approximately 20 min improved 20-min sprint times (quarters 2 and 4). Our study suggests that athletic performance such as sprinting and jumping are not altered by joint cooling applied prior to or during high-intensity intermittent exercise.

Urine liver-type fatty acid-binding protein predicts graft outcome up to 2 years after kidney transplantation.

BACKGROUND: Several new biomarkers for the detection of early tubular injury have been investigated in kidney transplant recipients. We recently identified day 2 urinary neutrophil gelatinase-associated lipocalin (NGAL) as a predictor of slow graft function and adverse 1-year outcome. In the present study, we further investigated the value of urinary NGAL and liver-type fatty acid binding protein (L-FABP) for predicting long-term graft outcomes up to 2 years. METHODS: This study was a single-center, prospective observational study. Serial urinary NGAL and L-FABP levels at 0 hours, 2 days, and 6 days after kidney transplantation (KT) were measured, and the clinical data were assessed during the 2-year period after KT. RESULTS: During the 2-year follow-up period, 13 (18.8%), 5 (7.2%), and 4 (5.8%) patients were diagnosed with acute T-cell-mediated rejection, acute antibody-mediated rejection (AMR) and chronic AMR, respectively. In addition, 10 patients (14.3%) developed calcineurin inhibitor toxicity and 6 (8.7%) developed BK viremia. The mean estimated glomerular filtration rates (eGFR) at 1 and 2 years after KT were 65.1 ± 19.1 and 58.5 ± 22.6 mL/min/1.73 m(2), respectively, When poor long-term graft function was defined as eGFR of less than 50 mL/min/1.73 m(2) at 2 years, elderly donors, acute rejection, and high 0-hour urinary L-FABP levels were significant risk factors. Furthermore, in rejection-free patients, L-FABP was strongly associated with poor long-term graft function (P = .006). Multivariate logistic regression analysis showed that high 0-hour L-FABP (P = .015) and acute rejection (P = .006) were independent factors predicting poor long-term graft function. Receiver operating characteristic analysis showed that the area under the curve for urinary L-FABP was 0.692 (P = .036). CONCLUSIONS: Our results suggest that urinary L-FABP may be a useful predictor of adverse long-term outcomes in KT patients.

Urine neutrophil gelatinase-associated lipocalin predicts graft outcome up to 1 year after kidney transplantation.

BACKGROUND: Several recent reports demonstrated the usefulness of new biomarkers in early prediction of delayed graft function (DGF) and graft recovery after kidney transplantation (KT). It is unknown, however, whether these biomarkers would predict long-term graft outcome. In this study, we examined whether the biomarkers including neutrophil gelatinase-associated lipocalin (NGAL) and liver-type fatty acid-binding protein (L-FABP) can predict 1-year graft outcome as well as short-term graft function especially in patients with early graft function (EGF). METHODS: This was a single-center, prospective observational study. Urine samples at 0 hours and 2 and 6 days were obtained and the level of NGAL and L-FABP were measured. RESULTS: Of the 69 KT recipients enrolled, seven developed DGF, and the remaining 62 patients were finally enrolled as EGF recipients. EGF recipients were additionally divided into immediate graft function (IGF, n = 48) and slow graft function (SGF, n = 14) groups. Urinary NGAL (u-NGAL) level on day 2, but not L-FABP nor serum creatinine, was significantly higher in SGF compared to IGF group. Higher day 2 u-NGAL level was associated with more frequent development of SGF and, in addition, with significantly lower 1-year estimated glomerular filtration rate (eGFR). In multivariate logistic regression analysis, day 2 u-NGAL was a significant, independent factor for predicting poor long-term graft function (1-year eGFR < 60 mL/min/1.73 m(2)). CONCLUSIONS: This study demonstrates the possibility that u-NGAL might be useful in predicting adverse 1-year outcome as well as short-term graft function even in EGF patients.

Choline transport via choline transporter-like protein 1 in conditionally immortalized rat syncytiotrophoblast cell lines TR-TBT.

Choline is an essential nutrient for phospholipids and acetylcholine biosynthesis in normal development of fetus. In the present study, we investigated the functional characteristics of choline transport system and inhibitory effect of cationic drugs on choline transport in rat conditionally immortalized syncytiotrophoblast cell line (TR-TBT). Choline transport was weakly Na(+) dependent and significantly influenced by extracellular pH and by membrane depolarization. The transport process of choline is saturable with Michaelis-Menten constants (K(m)) of 68microM and 130microM in TR-TBT 18d-1 and TR-TBT 18d-2 respectively. Choline uptake in the cells was inhibited by unlabeled choline and hemicholinium-3 as well as various organic cations including guanidine, amiloride and acetylcholine. However, the prototypical organic cation tetraethylammonium and cimetidine showed very little inhibitory effect of choline uptake in TR-TBT cells. RT-PCR revealed that choline transporter-like protein 1 (CTL1) and organic cation transporter 2 (OCT2) are expressed in TR-TBT cells. The transport properties of choline in TR-TBT cells were similar or identical to that of CTL1 but not OCT2. CTL1 was also detected in human placenta. In addition, several cationic drugs such as diphenhydramine and verapamil competitively inhibited choline uptake in TR-TBT 18d-1 with K(i) of 115microM and 55microM, respectively. Our results suggest that choline transport system, which has intermediate affinity and weakly Na(+) dependent, in TR-TBT seems to occur through a CTL1 and this system may have relevance with the uptake of pharmacologically important organic cation drugs.

The local prevalence of COPD by post-bronchodilator GOLD criteria in Korea.

OBJECTIVE: Chronic obstructive pulmonary disease (COPD) is a disease characterised by not fully reversible airflow limitation. The Global Initiative for Chronic Obstructive Lung Disease (GOLD) committee decided to diagnose COPD using post-bronchodilator spirometry values. We aimed to examine the prevalence and risk factors of COPD in Ansan, an industrialised city of Korea, by using the post-bronchodilator GOLD criteria. We then investigated the implications of brenchodilation on the prevalence of COPD. DESIGN: A total of 3642 participants in the Korean Health and Genome Study were interviewed about age, income, smoking status and respiratory symptoms and completed pulmonary function tests, including postbronchodilator spirometry. RESULTS: COPD prevalence by post-bronchodilator spirometry was 3.7% (134/3642), which was significantly different from that estimated using pre-bronchodilator criteria (7.7%, 282/3642). Exclusion of subjects with significant bronchodilator response (BDR) significantly lowered the prevalence of COPD to 3.3% (117/3572), compared with including subjects with post-bronchodilatory residual obstruction with significant BDR. Prevalence was associated with old age, smoking history, male sex and respiratory symptoms. CONCLUSION: COPD prevalence by post-bronchodilator GOLD criteria was 3.7%, which was much lower than that of pre-bronchodilator criteria. The bronchodilator reversibility test substantially affects estimations of COPD prevalence.

Inhibitory effects of Rumex japonicus Houtt. on the development of atopic dermatitis-like skin lesions in NC/Nga mice.

BACKGROUND: Rumex japonicus Houtt. (RJH) is one of the herbs used in Eastern countries for the treatment of atopic dermatitis (AD). It has been shown to have an antioxidative effect in human skin disease. OBJECTIVES: To examine whether RJH extract (RJH-E) suppresses the development of AD-like skin lesions in NC/Nga mice, which are induced by the repeated application of picryl chloride (PC). METHODS: The efficacy of RJH-E in NC/Nga mice was assessed by measuring symptom severity, scratching behaviour, Staphylococcus aureus numbers on an ear, and serum levels of IgE, interleukin (IL)-4 and interferon (IFN)-gamma. RESULTS: Oral administration of RJH-E to NC/Nga mice treated with PC inhibited the development of AD-like skin lesions as exemplified by a significant decrease in total skin symptom severity scores, and a decrease in hypertrophy, hyperkeratosis and infiltration of inflammatory cells in the skin. The scratching behaviour and numbers of S. aureus, which are known to be exacerbated in AD, were also significantly reduced by RJH-E. No significant change was observed in the serum levels of IFN-gamma, whereas IgE and IL-4 levels were significantly reduced by RJH-E. CONCLUSIONS: These results suggest that RJH-E inhibits the development of AD-like skin lesions in NC/Nga mice by suppressing the T-helper 2 cell response. Our results indicate that RJH treatment could provide an effective alternative therapy for the management of AD.

Visualization of bubble-fluid interaction by a moving object flow image analyzer system.

This paper deals with interaction between a bubble and fluid around it, visualized by a moving object flow image analyzer (MOFIA) consisting of a three-dimensional (3D) moving object image analyzer (MOIA) and two-dimensional particle image velocimetry (PIV). The experiments were carried out for rising bubbles of various sizes and shapes in stagnant water in a vertical pipe. In the MOFIA employed, 3D-MOIA was used to measure bubble motion and PIV to measure fluid flow. The 3D position and shape of a bubble and the velocity field were measured simultaneously. The experimental results showed that the interaction was characterized by the shape, size and density of a bubble. Concretely, they showed the characteristics of bubble motion, wake shedding, and flow field.

Accuracy of reformatted CT image for measuring the pre-implant site: analysis of the image distortion related to the gantry angle change.

OBJECTIVES: In a multiplanar reconstruction CT program (CT/MPR), the altered gantry angle corresponding to the patient's position during scanning for multiplanar reformatted CT may cause distortion of the image. The aim of this study was to quantitatively assess the distortion in reformatted central panoramic and cross-sectional images owing to the change of gantry angle. METHODS: A resin block model with four cylindrical holes and a human dry mandible were used in high resolution conventional CT scanning. A MPR software package was used for reformatted panoramic and cross-sectional images. The block and the gantry were equally inclined at 0 degrees, 15 degrees and 30 degrees. The relationship between the positional difference and depth of the hole as determined in the reformatted image, and the actual depth was analysed with respect to the gantry angle. The actual depths (H) were calculated by use of the measured depths (h) and the inclined angles: H=hxcos beta. The positional differences of the hole bases (W) were also calculated by use of the measured depths and the inclined angles: W=hxsin beta or Hxtan beta. RESULTS: Increasing the gantry angle, increased the depth and inclination of the holes in the reformatted central panoramic images. Inclined angle (beta) was the same as gantry angle. CONCLUSION: When the gantry angle is not at 0 degrees, certain CT/MPR programs may distort the reformatted image. If distortion occurs, the corrected position and usable length of pre-implant sites can be calculated by use of the above formula.

Quantitative analysis of normal major salivary glands using computed tomography.

OBJECTIVES: We sought to calculate the size and the computed tomography (CT) number of normal parotid and submandibular glands and to evaluate their relationship with respect to sex, age, and obesity in the Korean population. STUDY DESIGN: The authors investigated the axial CT images of 42 healthy volunteers. The maximum cross-sectional area (MCSA) was used as an indicator of the size of the gland. Three regions of interest on axial scans were selected to calculate the mean CT number. RESULTS: There was a significant decrease in the mean MCSA of the submandibular gland and the CT number of both glands with age. The MCSA of the submandibular gland in males was larger than that in females. There was a close correlation between the parotid and the submandibular glands with respect to the CT number, as well as between the left and right glands with respect to the MCSA and the CT number. The body mass index was positively correlated with the MCSA of the parotid gland, whereas the body mass indexes and the CT numbers of both glands were negatively correlated. CONCLUSIONS: Both age and obesity are closely correlated with the size and the CT number of the major salivary glands. Moreover, the correlation between the CT numbers of the parotid and the submandibular glands may be used for diagnostic purposes.

Quantitative analysis of apical root resorption by means of digital subtraction radiography.

OBJECTIVE: This study was performed to assess the diagnostic and quantifying ability of digital subtraction images for simulated apical root resorption, as well as to compare the diagnostic accuracy of conventional intraoral radiographs with digital subtraction images for this condition. STUDY DESIGN: Digital and intraoral radiographs of 10 sound maxillary central incisors and those with simulated apical root resorption were taken with varying horizontal and vertical angulations of the x-ray beam. Paired t tests were used to compare the lengths of the sound teeth on the images with their actual lengths, and the estimated amount of simulated apical root resorption was compared with the actual amount of tooth loss by means of Emago software. The diagnostic accuracy for detecting lesions was also evaluated on conventional intraoral radiographs and digital subtraction images through receiver operating characteristic (ROC) analysis. RESULTS: There were no statistically significant differences between the actual lengths and those measured on the reconstructed images of the sound teeth. The calculated amounts of apical root resorption showed no statistically significant differences in comparison with the actual amounts (P >.05). The diagnostic accuracy of the conventional intraoral radiographs in detecting the lesions was low (ROC area = 0.6446). CONCLUSION: A quantitative analysis of small amounts of apical root resorption can be performed by means of digital subtraction radiography.

Transcriptional repression of type I procollagen genes during adipocyte differentiation.

Adipocyte differentiation is a very complex process in which whole-cell changes are accompanied. Among them, type I procollagen gene has been shown to specifically decrease during adipocyte differentiation; however, little is known about the molecular mechanism. To examine how type I procollagen gene expression is regulated at the level of transcription during adipocyte differentiation, 3T3-L1 preadipocyte cell line was used as an in vitro model. Northern blot analysis demonstrated that mRNA expression of type I procollagen gene was dramatically reduced during adipocyte differentiation. Time-course analysis indicated that decrease in mRNA expression occurred at early stage of differentiation. Studies on several stable cell lines showed that transcriptional activities of both alpha1 and alpha2 promoters decreased significantly during adipocyte differentiation. Despite extensive deletion-promoter analyses, however, we could not identify the cis-element responsible for the switch-off of type I procollagen gene during adipocyte differentiation, suggesting that the transcriptional repression of this gene occur through general transcription machinery rather than a specific cis-element. In conclusion, down-regulation of type I procollagen mRNA expression during adipocyte differentiation is due to repression of its promoter activity through general transcription machinery.

Clinical significance of early diastolic notch depth: uterine artery Doppler velocimetry in the third trimester.

OBJECTIVE: Our purpose was to evaluate the usefulness of early diastolic notch depth in predicting adverse perinatal outcome. STUDY DESIGN: Uterine artery Doppler velocimetry is widely applied in predicting pregnancy outcomes. Recent studies of uterine artery early diastolic notch have demonstrated its usefulness as a marker for fetal well-being. The early diastolic notch represents the reflected blood flow of uteroplacental circulation. This study was carried out under the hypothesis that when early diastolic notch is present evaluating its depth plays a significant role in predicting poor pregnancy outcomes. Its ability to predict adverse perinatal outcomes was evaluated in 198 pregnant women with early diastolic notch after 28 weeks' gestation. RESULTS: The incidence of adverse perinatal outcome (5-minute Apgar score <7, cesarean delivery because of fetal distress, admission to neonatal intensive care unit, fetal growth restriction, or perinatal death) was 90.5% when the notch index was /=0.90, the respective incidences were 72.3%, 46. 9%, and 28.1%. CONCLUSION: This study suggests that evaluating notch depth in the presence of early diastolic notch on uterine artery velocimetry during the third trimester would be a useful method for predicting adverse perinatal outcomes.

Combined cis-platinum and alpha interferon therapy of advanced hepatocellular carcinoma.

To evaluate the clinical efficacy of alpha-interferon(IFN-alpha) plus cis-platinum in hepatocellular carcinoma(HCC). 56 inoperable patients with HCC were divided into IFN-alpha plus cis-platinum treated group (n = 30) and no antitumor therapy group (n = 26). The survival of IFN-alpha plus cis-platinum treated patients was significantly better than that of patients who received no antitumor therapy (p = 0.001). Median survival time was 33 weeks and 14.0 weeks, respectively. The cumulative estimated survival rates of our IFN-alpha plus cis-platinum treated group (93.5% at 3mo, 75.0% at 6mo) were for longer than that of the no antitumor therapy group (84.6% at 3mo, 57.7% at 6mo). Objective tumor regression, greater than 50% was observed in 13.3% (4 of 30) of patients receiving IFN-alpha plus cis-platinum. By the univariate analysis, the absence of portal vein thrombus (p < 0.05), alkaline phosphatase lesser than 280 U/L (p = 0.001), total bilirubin less than 2.0 mg% (p < 0.05), serum triglyceride less than 155 mg/dl (p < 0.05) were shown to be the factors most significantly favoring a better survival. By the multivariate analysis, using Cox proportional hazards model, IFN-alpha plus cis-platinum treated group (p = 0.0001), alkaline phosphatase less than 280 mg/dl (p = 0.005), the absence of portal vein thrombus (p = 0.020) were independent favorable prognostic factors. We conclude that IFN-alpha plus cis-platinum is useful in patients with inoperable HCC and the above favorable prognostic factors may also be useful in the design and analysis of future clinical trials of systemic chemotherapy for HCC.

Oil sorption behavior of various sorbents studied by sorption capacity measurement and environmental scanning electron microscopy.

Oil sorption capacities of various natural and man-made fibrous sorbents were compared in a simulated seawater bath containing oil. Natural sorbents such as milkweed, kapok, cotton, and wool showed higher sorption capacities than man-made sorbents such as polyester, polypropylene, viscose rayon, nylon 6, nylon 66, and acetate. Sorption capacities of the natural sorbents were over 30 g oil/g fiber. No definite advantages were observed using man-made bicomponent and biconstituent fibers over regular man-made fibers with respect to their sorption capacity. Analyses of sorption mechanisms using an environmental scanning electron microscope revealed that an oil deposit disappeared from the fiber surface after a certain time interval in milkweed, kapok, and cotton. This suggested that the sorption of oil in these fibers occurred through capillary action, probably due to their hollow lumens. Contrarily, adsorption, a surface phenomenon, would be the most prominent mechanism for oil sorption of wool fibers due to large amounts of surface wax, irregular scaly surfaces, and crimp. Effects of both adsorption and absorption were shown in the oil sorption of man-made fibers, depending upon the type and shape of the sorbent. Dumbbell-like oil deposits were seen on the fiber surface in certain oleophilic man-made fibers, because of a partial wetting of oil on the fiber surface. For some hydrophilic man-made fibers such as polyvinylalcohol and copolymer of isobutylene-maleic anhydride, the physical configuration of the fiber was a decisive factor in determining oil sorpton capacity of the sorbents.

Cutaneous metastatic adenocarcinoma of stomach--nodular and inflammatory carcinoma.

A 65-year-old male patient presented multiple cutaneous nodules on trunk and an erythematous inflammatory lesion resembling erysipelas on left side of neck. Result of skin biopsies disclosed groups of metastatic adenocarcinoma cells in the dermis, subcutaneous tissue, and lumina of the lymphatic vessels. Histopathologic examination of the stomach, along with the gastroscope, established that the cutaneous metastasis was of stomach origin.